Effect of ozone versus naringin on testicular injury in experimentally induced ulcerative colitis in adult male albino rats.

Testicular dysfunction is caused by the continuous inflammation and oxidative stress that are present at the local site in ulcerative colitis (UC) spreading to the testes via systemic circulation. The influence of ozone and naringine on colitis-mediated testicular dysfunction was investigated in this study. Forty-eight adult male rats were divided into four groups: I control group, II dextran sodium sulfate (DSS) UC-induced group, III DSS+naringine, and IV DSS+ozone groups. UC was induced in groups II, III, and IV using 0.1 ml of 4% DSS in their drinking water per day for 6 days by gastric gavage. All animals were sacrificed 45 days from the start. Blood samples were obtained to estimate serum testosterone hormone. Testicular tissues were processed for measurement of tissue malondialdehyde (MDA) and examined by light and electron microscopes. Ultrastructurally, group II revealed a relatively thick basement membrane enveloping the seminiferous tubule. Sertoli cell cytoplasm appears rarified with wide intracellular spaces, vacuoles, and multiple lysosomes; distorted spermatogonia with electron dense nuclei and cytoplasm; and primary spermatocytes with small nuclei and electron dense cytoplasm. Abnormal sperm profiles were visible in middle pieces, mid, principle, and end pieces that were markedly affected with disorganization of axoneme and outer dense fibers. Leydig cells revealed dilated cisternae of smooth endoplasmic reticulum. Morphometric and statistical analyses were performed. Group III showed some improvement; however, group IV showed more improvement. The results indicated that ozone caused marked improvement than naringine against UC-induced testicular damage via their antioxidant and anti-inflammatory properties.

Oral supramolecular nanovectors for dual natural medicine codelivery to prevent gastric mucosal lesion.

The oral administration of a single formulation loaded with more than one natural medicine to treat chronic diseases has advantages such as convenience, effectiveness, and economy. Here, using biomaterials approved by the drug administration, we fabricated supramolecular nanovectors containing dual natural medicines to prevent gastric mucosal lesions. Nanovectors exhibited superior intestinal absorption and bioavailability, which might be due to their high dispersion, good muco-adhesiveness, blood-lymph circulation transport, lipid sensing, and protective effects. Molecular docking results clarified the possible mechanisms in aspects of efflux pump (p-glycoprotein and multidrug resistance protein 1) inhibition effects, metabolic enzyme (cytochrome P450 3A4/1A2) blocking effects, serum albumin deposit effects, and dual drug interaction effects. Nanovectors decreased ethanol-induced gastric mucosal lesions by lowering the gastric ulcer index, preventing oxidative damage, decreasing interleukin-6, tumor necrosis factor-α and malondialdehyde, increasing glutathione, superoxide dismutase, and prostaglandin E2 levels. The interactions of inhibitor of nuclear factor-κB or κB kinase-related proteins and dual drugs or nanovector components were simulated computationally to provide an understanding of the gastro-protective action mechanism. In all, industrializable supramolecular nanovectors could effectively co-deliver dual natural medicines via the oral route by improving the pharmacokinetic behavior and exerting protective efficacy of the gastric mucosa by decreasing the oxidative stress and inflammatory level.

[Effect of melatonin on p38MAPKsignaling pathway in rats with phosgene-induced lung injury].

OBJECTIVE: To investigate the effect of melatonin (MT) on p38 mitogen-activated protein kinase (MAPK) signaling pathway in rats with phosgene-induced lung injury. METHODS: Fifty specific pathogen-free male Sprague-Dawley rats were randomly divided into phosgene inhalation group, air control group, saline control group, MT treatment group, and SB203580 (specific inhibitor of p38 MAPK) group, with 10 mice in each group. All groups except the air control group were exposed to phosgene, and the animals were sacrificed 6 h later. Lung wet/dry weight (W/D) ratio and the content of malondialdehyde (MDA) and nitric oxide (NO) and activity of myeloperoxidase (MPO) in bronchoalveolar lavage fluid (BALF) were measured. The qualitative and quantitative expression of p38 MAPK and phospho-p38 MAPK (p-p38) was measured by immunohistochemistry (IHC) and Western blot, respectively. Inducible nitric oxide synthase (iNOS) level in lung tissue was determined by Western blot. RESULTS: Compared with the air control group, the phosgene inhalation group had significantly increased lung W/D ratio and neutrophil count in BALF (P < 0.01); the MT treatment group had significantly lower neutrophil count and lung W/D ratio than the phosgene inhalation group (P < 0.05). IHC demonstrated that the air control group had relatively weak expression of p-p38 in lung tissue; the expression of p-p38 was significantly up-regulated after phosgene inhalation, and it was mainly distributed in infiltrating inflammatory cells and vascular endothelial cells, positive in the cytoplasm and nucleus of many cells. The distribution of p-p38-positive cells in the MT treatment and SB203580 groups was similar to that in the phosgene inhalation group, but the MT treatment and SB203580 groups had a significantly reduced number of cells with p-p38-positive nuclei and a significantly reduced intensity of p-p38 expression signals. The phosgene inhalation group had significantly increased content of MDA and NO and activity of MPO compared with the air control group (P < 0.01); the MT treatment and SB203580 groups had significantly reduced content of MDA and NO and activity of MPO compared with the phosgene inhalation group (P < 0.05), but had higher content of MDA and NO and activity of MPO than the air control group. The Western blot showed that the phosgene inhalation group had significantly increased expression of iNOS and p-p38 compared with the air control group (P < 0.01); the MT treatment and SB203580 groups had lower expression of iNOS and p-p38 than the phosgene inhalation group (P < 0.05). CONCLUSION: MT and SB203580 have a significant protective effect in rats with phosgene-induced lung injury, and the mechanism may be associated with scavenging free radicals and inhibiting activation of p38 MAPK and expression of iNOS.

Micronuclei in cord blood lymphocytes and associations with biomarkers of exposure to carcinogens and hormonally active factors, gene polymorphisms, and gene expression: the NewGeneris cohort.

BACKGROUND: Leukemia incidence has increased in recent decades among European children, suggesting that early-life environmental exposures play an important role in disease development. OBJECTIVES: We investigated the hypothesis that childhood susceptibility may increase as a result of in utero exposure to carcinogens and hormonally acting factors. Using cord blood samples from the NewGeneris cohort, we examined associations between a range of biomarkers of carcinogen exposure and hormonally acting factors with micronuclei (MN) frequency as a proxy measure of cancer risk. Associations with gene expression and genotype were also explored. METHODS: DNA and protein adducts, gene expression profiles, circulating hormonally acting factors, and GWAS (genome-wide association study) data were investigated in relation to genomic damage measured by MN frequency in lymphocytes from 623 newborns enrolled between 2006 and 2010 across Europe. RESULTS: Malondialdehyde DNA adducts (M1dG) were associated with increased MN frequency in binucleated lymphocytes (MNBN), and exposure to androgenic, estrogenic, and dioxin-like compounds was associated with MN frequency in mononucleated lymphocytes (MNMONO), although no monotonic exposure-outcome relationship was observed. Lower frequencies of MNBN were associated with a 1-unit increase expression of PDCD11, LATS2, TRIM13, CD28, SMC1A, IL7R, and NIPBL genes. Gene expression was significantly higher in association with the highest versus lowest category of bulky and M1dG-DNA adducts for five and six genes, respectively. Gene expression levels were significantly lower for 11 genes in association with the highest versus lowest category of plasma AR CALUX® (chemically activated luciferase expression for androgens) (8 genes), ERα CALUX® (for estrogens) (2 genes), and DR CALUX® (for dioxins). Several SNPs (single-nucleotide polymorphisms) on chromosome 11 near FOLH1 significantly modified associations between androgen activity and MNBN frequency. Polymorphisms in EPHX1/2 and CYP2E1 were associated with MNBN. CONCLUSION: We measured in utero exposure to selected environmental carcinogens and circulating hormonally acting factors and detected associations with MN frequency in newborns circulating T lymphocytes. The results highlight mechanisms that may contribute to carcinogen-induced leukemia and require further research.

Estrés oxidativo en cefalópodos: I. Determinación de TBARS / Oxidative stress in cephalopods: I. TBARS determination

El estrés oxidativo se produce cuando se genera un desbalance desfavorable entre las especies reactivas del oxígeno y las defensas antioxidantes, provocando daño oxidativo a macromoléculas. Varios estudios han resaltado la importancia del estrés oxidativo en el campo de la ecotoxicología, particularmente su relación con el impacto que generan los contaminantes que alcanzan los cuerpos de agua. El cuantifcar los parámetros de estrés oxidativo ha permitido el uso de los mismos como herramienta de diagnóstico (biomarcadores), con capacidad predictiva del impacto de los contaminantes sobre los organismos. Uno de los índices más frecuentemente utilizados para estimar el daño oxidativo a lípidos es la determinación de sustancias reactivas al ácido tiobarbitúrico (TBARS), producto fnal de la peroxidación lipídica. Octopus tehuelchus es un importante recurso pesquero en la costa patagónica, expuesto en algunas áreas a contaminación antrópica. Dado que el estudio de parámetros de estrés oxidativo aún no ha sido abordado en esta Clase de moluscos y que en muchos modelos biológicos, los contaminantes ambientales actúan generando estrés oxidativo, es clave encontrar sus blancos de acción, para empezar a caracterizar las alteraciones metabólicas y fsiológicas asociadas a su mecanismo de acción. El objetivo de este trabajo fue la puesta a punto del método de determinación de daño oxidativo a lípidos en distintos tejidos del pulpo Octopus tehuelchus desde modelos previamente ensayados en el laboratorio.

Oxidative stress occurs when there is an unfavorable imbalance between reactive oxygen species and antioxidant defenses, causing oxidative damage to macromolecules. Several studies have highlighted the importance of oxidative stress in the ecology feld related to the impact generated by pollutants reaching water bodies. The quantifcation of oxidative stress parameters led to their use as diagnostic tools (biomarkers) with predictive capability of showing the impact of pollutants on organisms. One of the most frequently used indexes to estimate the oxidative damage to lipids is the determination of reactive thiobarbituric acid substances (TBARs) (fnal product of lipid peroxidation). Octopus tehuelchus is an important fshery resource in the Patagonian coast exposed to anthropogenic pollution. The study of oxidative stress parameters has not been yet tackled in this class of molluscs. Taking into account that, in many biological models, environmental pollutants generate oxidative stress, it is important to fnd their targets of action, to start to characterize metabolic and physiological alterations associated to their mechanisms of action. The aim of this work was to adjust the method of determination of oxidative damage to lipids in various tissues of the octopus, Octopus tehuelchus, from models previously tested in the laboratory.

Streptococci-human papilloma virus interaction with ethanol exposure leads to keratinocyte damage.

PURPOSE: Ethanol, human papilloma virus (HPV), and poor oral hygiene are risk factors that have been attributed to oral carcinogenesis. Streptococci sp and HPV infections are common in the head and neck, often associated with sexual activity. Although HPV is linked to head and neck squamous cell carcinoma, it is unclear whether there is a similar role for Streptococci sp. This cell study examines whether Streptococci sp and HPV-16 with exposure to ethyl alcohol (ETOH) can act as cofactors in the malignant transformation of oral keratinocytes. MATERIALS AND METHODS: ETOH (0.1%-20% vol/vol) was used to investigate Streptococci sp attachment with immortalized E6-expressing HPV/HOK-16B cells, human oral buccal keratinocytes, and foreskin keratinocytes. Streptococci sp (Streptococci mutans [LT11]) and various strains of acetaldehyde (AA) producer and nonproducer Streptococcus salivarius (110-1, 109-2, 101-7, and 107-1) and a lactic acid producer bacterium, Lactobacillus rhamnosus (24-1 and 25-2), were examined for interactions with keratinocytes by use of a green dye (percent of cells with colonies after 24 hours). Carcinogens, AA, malondialdehyde, DNA damage, and proliferation (5'-bromo-2-deoxyuridine) among keratinocytes were also quantified. RESULTS: AA and malondialdehyde production from permissible Streptococci sp significantly increased with attachment to keratinocytes, whereas L rhamnosus did not significantly attach to keratinocytes. This attachment was associated with enhanced levels of AA adduct formation, proliferation (5'-bromo-2-deoxyuridine incorporation), and enhanced migration through integrin-coated basement membrane by HPV oral keratinocytes, which are characteristics of a malignant phenotype. CONCLUSIONS: These cell studies suggest that oral Streptococci sp and HPV (HPV-16) cooperate to transform oral keratinocytes after low-level ETOH (1%) exposure. These results appear to suggest a significant clinical interaction, but further validation is warranted.

Poly(ADP-ribose) polymerase inhibition modulates experimental acute necrotizing pancreatitis-induced oxidative stress, bacterial translocation and neopterin concentrations in rats.

Various studies have been performed to find out novel treatment strategies for acute necrotizing pancreatitis (ANP). Inhibition of poly(ADP-ribose) polymerase (PARP) is shown to reduce inflammation in several pathological conditions. We aimed to evaluate the efficacy of benzamide, a PARP inhibitor, in an experimental model of ANP. Thirty Sprague-Dawley rats were divided into three groups: sham-operated, ANP and ANP + benzamide groups. All groups except the sham-operated group were subjected to the ANP procedure, induced by infusing of 1 mL/kg of 3% sodium taurocholate into the common biliopancreatic duct. The ANP + benzamide group received 100 mg/kg/day benzamide intraperitoneally for a total of three days after induction of pancreatitis. The surviving animals were killed at the fourth day and the pancreas was harvested for biochemical, microbiological and histological analysis. Blood samples were also obtained from the animals. In the ANP group, a significant increase was observed in concentrations of serum amylase and neopterin and tissue oxidative stress indices (malondialdehyde, superoxide dismutase and glutathione peroxidase). Almost all of these changes were found to be reversed to near their normal values in the ANP + benzamide group. Histological injury scores were significantly higher in the ANP group than in the sham group (P < 0.05, ANP versus sham), and were significantly lower in the ANP + benzamide group than in the ANP group (P < 0.05, ANP + benzamide versus ANP). Evaluation of bacterial translocation identified significantly fewer infected sites in the ANP + benzamide group than in the ANP animals (P < 0.01). We observed that inhibition of PARP with benzamide reduced the severity, the mortality, the bacterial translocation rates and the neopterin concentrations in an experimental ANP model in rats. These findings suggest that it may be possible to improve the outcome of ANP by using PARP inhibitors.

Lipid peroxidation-DNA damage by malondialdehyde.

Malondialdehyde is a naturally occurring product of lipid peroxidation and prostaglandin biosynthesis that is mutagenic and carcinogenic. It reacts with DNA to form adducts to deoxyguanosine and deoxyadenosine. The major adduct to DNA is a pyrimidopurinone called M1G. Site-specific mutagenesis experiments indicate that M1G is mutagenic in bacteria and is repaired by the nucleotide excision repair pathway. M1G has been detected in liver, white blood cells, pancreas, and breast from healthy human beings at levels ranging from 1-120 per 108 nucleotides. Several different assays for M1G have been described that are based on mass spectrometry, 32P-postlabeling, or immunochemical techniques. Each technique offers advantages and disadvantages based on a combination of sensitivity and specificity. Application of each of these techniques to the analysis of M1G is reviewed and future needs for improvements are identified. M1G appears to be a major endogenous DNA adduct in human beings that may contribute significantly to cancer linked to lifestyle and dietary factors. High throughput methods for its detection and quantitation will be extremely useful for screening large populations.

Oral toxicity of malonaldehyde: a 90-day study on mice.

The oral toxicity of malonaldehyde (MA), a product of lipid peroxidation found in some foods, was investigated in a 90-d study on mice, MA as the sodium enol salt was administered in the drinking water to 8-wk-old female Swiss mice at levels calculated to provide 2, 10, 50, 250, or 500 micrograms/g body weight . d. There was no mortality and all groups gained weight at comparable rates except that those that received 500 micrograms/g body weight . d gained more slowly and lost weight after 50 d. Histopathological examination of 27 tissues indicated that the liver was the only organ that underwent dose-dependent changes. All levels of MA induced irregularities (anisokaryosis, hyperchromicity, vesiculation) of hepatic nuclei. Pancreatic lesions consisting primarily of atrophy of the exocrine cells with loss of zymogen granulation occurred in animals which received 500 micrograms MA/g body weight . d. Mild dysplasia of the urinary bladder epithelium was found in all treatment groups. Approximately 1% of the dose was excreted unchanged in the urine at each level of administration.

Protective effect of riboflavin on suppression of growth caused by oxidized oil.

When oxidized corn oil (100 nmol in terms of malondialdehyde/day/rat) was administered to a riboflavin-deficient rat, the body weight gain was markedly suppressed. However, when 20 microgram of riboflavin/day/rat was administered with the oxidized corn oil, reasonable growth and normal flavin levels in the liver, kidney and heart could be attained, though they were somewhat less than those of the animals fed on the normal diet containing non-oxidized corn oil. It was noted that the elevation of lipid peroxide level in blood plasma of animals administered with the oxidized oil was effectively prevented by riboflavin. These results indicate the protective effect of riboflavin on suppression of growth caused by the oxidized oil.